Mechanisms of DNA uptake by mammalian cells: Fate of exogenously added DNA monitored by the use of fluorescent dyes

A. Loyter, G. A. Scangos, F. H. Ruddle

Research output: Contribution to journalArticlepeer-review

190 Scopus citations

Abstract

Coprecipitation of DNA with calcium phosphate is a commonly used method of gene transfer in mammalian cells. We have found that DNA forms a tight complex with Ca P[i] and that DNA in this complex is resistant to nucleases present in serum or added externally. Under optimal conditions, virtually all of the recipient mouse Ltk-Aprt- cells take up Ca P[i]-DNA complexes, as determined by fluorescent dyes specific for DNA (4',6-diaminilo-2-phenylindol dihydrochloride) or for calcium salts (chlorotetracycline). However, only a small proportion of the cells have detectable Ca P[i]-DNA complexes in the nucleus. Uptake of the CaP[i]-DNA complexes was highly dependent upon the pH at which the CaP[i]-DNA complex was formed and upon the concentration of DNA in the complex.

Original languageEnglish
Pages (from-to)422-426
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume79
Issue number2 I
DOIs
StatePublished - 1982
Externally publishedYes

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